Hindawi Evidence-Based Complementary and Alternative Medicine Volume 2021, Article ID 6622894, 7 pages https://doi.org/10.1155/2021/6622894

Research Article Antiprotozoal Compounds from Urolepis hecatantha ()

Orlando G. Elso ,1,2 Maria Clavin ,1,2 Natalia Hernandez ,1 Toma´s Sgarlata ,1 Herna´n Bach ,3,4 Ce´sar A. N. Catalan ,5 Elena Aguilera ,6 Guzman Alvarez ,7 and Valeria P. Su¨lsen 1,2

1Universidad de Buenos Aires, Facultad de Farmacia y Bioqu´ımica, Ca´tedra de Farmacognosia, Jun´ın 956, Buenos Aires 1113, 2CONICET–Universidad de Buenos Aires, Instituto de Qu´ımica y Metabolismo del Fa´rmaco (IQUIMEFA), Jun´ın 956, Buenos Aires 1113, Argentina 3Universidad de Buenos Aires, Facultad de Farmacia y Bioqu´ımica, Museo de Farmacobot´anica, Jun´ın 956, Buenos Aires 1113, Argentina 4Instituto de Recursos Biol´ogicos INTA-Hurlingham, De los Reseros y N. Repetto (1686), Hurlingham, Buenos Aires, Argentina 5Universidad Nacional de Tucuma´n, Facultad de Bioqu´ımica Qu´ımica y Farmacia, Instituto de Qu´ımica Orga´nica, Ayacucho 471 (T4000INI), San Miguel de Tucuma´n, Argentina 6Grupo de Qu´ımica Medicinal-Laboratorio de Qu´ımica Org´anica, Facultad de Ciencias, Universidad de la Rep´ublica, Montevideo 11400, 7Laboratorio de Mole´culas Bioactivas, Universidad de la Repu´blica, CENUR Litoral Norte, Paysandu´ 60000, Uruguay

Correspondence should be addressed to Valeria P. S¨ulsen; vsulsen@ffyb.uba.ar

Received 20 October 2020; Revised 1 February 2021; Accepted 3 February 2021; Published 12 February 2021

Academic Editor: Shagufta Perveen

Copyright © 2021 Orlando G. Elso et al. *is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. *e dewaxed dichloromethane extract of Urolepis hecatantha and the compounds isolated from it were tested for their in vitro activity on Trypanosoma cruzi epimastigotes and Leishmania infantum promastigotes. *e extract of U. hecatantha showed activity against both parasites with IC50 values of 7 µg/mL and 31 µg/mL, respectively. Fractionation of the dichloromethane extract led to the isolation of euparin, jaceidin, santhemoidin C, and eucannabinolide. *e sesquiterpene lactones eucannabinolide and santhemoidin C were active on T. cruzi with IC50 values of 10 ± 2 µM (4.2 µg/mL) and 18 ± 3 µM (7.6 µg/mL), respectively. Euparin and santhemoidin C were the most active on L. infantum with IC50 values of 18 ± 4 µM (3.9 µg/mL) and 19 ± 4 µM (8.0 µg/ mL), respectively. Eucannabinolide has also shown drug-like pharmacokinetic and toxicity properties.

1. Introduction attributed to this parasitosis [2]. Chagas’ disease was his- torically linked to poor rural areas of Latin America where Chagas’ disease and leishmaniasis are protozoan parasitic the insect vector is present. In recent years, the disease has diseases caused by Trypanosoma cruzi and different species spread to cities and nonendemic areas due to migrations of of the genus Leishmania and transmitted by infected blood- infected people and nonvectorial transmission of the par- sucking triatomine bugs and phlebotomine sandflies, re- asite, turning Chagas’ disease into a global public health spectively. *ey are both classified as neglected tropical problem [3]. Nifurtimox and benznidazole are the only diseases by the World Health Organization (WHO) [1]. drugs currently available for Chagas’ disease treatment. Both Chagas’ disease or American Trypanosomiasis is a po- drugs are effective in the acute stage of the infection and tentially life-threatening disease that affects 6 to 7 million vertical transmission prevention, but their efficacy dimin- people worldwide. It is estimated that about 30000 new cases ishes in the chronic phase. Besides, frequent adverse events occur annually and more than 12000 deaths per year are lead to high rates of treatment discontinuation [4]. 2 Evidence-Based Complementary and Alternative Medicine

*erefore, the development of new trypanocidal drugs for dichloromethane (3 × 60 mL). Dichloromethane subextracts Chagas’ disease treatment is needed. were joined and taken to dryness on a rotary evaporator to Leishmaniasis has three clinical forms: cutaneous, mu- yield 15 g of dewaxed extract. Dewaxed extract (DDE) was cocutaneous, and visceral also known as kala-azar. Although fractionated by silica gel column chromatography cutaneous leishmaniasis is the most common form, visceral (60 × 5 cm, 220 g, 230–400 mesh) and eluted with a gradient leishmaniasis is the most severe form [5]. According to the of dichloromethane (CH2Cl2) and increasing amounts of WHO, more than one billion people are at risk of infection. ethyl acetate (EtOAc): 100% CH2Cl2, CH2Cl2: EtOAc (9 :1), It is estimated that 30000 new cases of visceral leishmaniasis (8 : 2), (7 : 3), (6 : 4), (5 : 5), (4 : 6), (3 : 7), (2 : 8), (1 : 9), and and more than one million new cases of cutaneous leish- 100% EtOAc. Fractions (5 × 200 mL) of each solvent ratio maniasis occur annually [5]. *e chemotherapy of leish- (A1−5 to K1−5) were collected. All column chromatography maniasis is based on the use of sodium stibogluconate, fractions were monitored by thin-layer chromatography meglumine antimoniate, pentamidine, amphotericin B, using silica gel 60 F254 plates and anisaldehyde sulphuric as paromomycin, and miltefosine. *ese drugs are toxic and spraying reagent. have other limitations such as the route of administration, length and cost of treatment, and emergence of drug re- sistance [6]. 2.3. Compounds Isolation. Fractions A5 (100% DCM) and Natural products play an important role in the drug B1−5 (DCM: EtOAc 9: 1) were pooled and the solvent was discovery process. One of the most relevant examples is evaporated on a rotary evaporator. *e residue obtained was artemisinin, a sesquiterpene lactone isolated from Artemisia transferred to a small vial with a minimum amount of ethyl annua currently used for malaria treatment [7]. Several acetate and the solution was left at room temperature natural products with promissory activity against patho- overnight. From this solution, yellow acicular crystals of genic protozoa have been reported [8, 9]. compound 1 (euparin, 28 mg) were obtained. Urolepis hecatantha (DC) R. M. King and H. Rob. (syn. Fractions D2−5, eluted with CH2Cl2 : EtOAc (7 : 3), Eupatorium hecatanthum (DC) Baker) is the only species of and the fraction E1, eluted with CH2Cl2 : EtOAc (6 : 4), the monotypic genus Urolepis (Asteraceae) [10]. *e eth- showing a similar profile on TLC, were reunited and nomedical uses of the aerial parts of U. hecatantha by in- brought to dryness on a rotary evaporator. *e residue digenous groups of northeast Argentina have been reported was suspended in a minimum amount of dichloro- [11–13]. *e fresh aerial parts are chewed as antitussive [11], methane and purified by preparative TLC using silica gel while the infusion or decoction of the aerial parts is used plates. *e plates were developed using toluene: EtOAc : topically for gangrene and ulceration treatment [12]. *is formic acid (6 : 4 :1) as a mobile phase. After drying, species has been employed also as an analgesic for teeth pain plates were analyzed under UV light where a deep green treatment [13]. *e isolation of flavonoids, terpenoids, and fluorescent band (Rf � 0.6) was observed. *e fluorescent benzofuran derivatives from a collection of U. hecatantha band was scraped out and extracted with methanol. After from Bolivia has been reported [14]. solvent evaporation, a yellow powder (10 mg) identified In this work, we report the isolation of four compounds as jaceidin was obtained. from U. hecatantha from Argentina and the evaluation of Fractions H2-4, eluted with CH2Cl2 : EtOAc (3 : 7), their in vitro activity on Trypanosoma cruzi and Leishmania were pooled and concentrated under vacuum in a rotary infantum. *e toxicity and pharmacokinetic properties of evaporator. *e residue was dissolved with a minimum the compounds were also estimated. amount of ethyl acetate and allowed to stand at room temperature for 24 hours. From this solution, pure 2. Materials and Methods crystals of compound 3 (santhemoidin C, 120 mg) were obtained. Fractions I3-5, eluted with CH2Cl2 : EtOAc (2 : 2.1. Materials. *e aerial parts of U. hecatantha (DC.) 8), and fractions J1-3, eluted with CH2Cl2 : EtOAc (1 : 9), R. King and H. Robins (Asteraceae) were collected in Buenos were combined and brought to dryness on a rotary Aires province, Argentina, in March 2018. *e plant material evaporator. *e residue was fractionated by silica gel was identified and deposited at the Herbarium of the Faculty column chromatography (50 × 3 cm, 150 g, 230–400 mesh) of Pharmacy and Biochemistry, University of Buenos Aires and eluted isocratically with a 2 :1 mixture of CH2Cl2 : (BAF 16100). EtOAc. Twenty fractions of 50 mL each were collected. Fractions 15–17 were reunited and brought to dryness on a rotary evaporator to afford compound 4 (eucannabi- 2.2. Extraction and Fractionation Procedures. Grounded nolide, 108 mg) as a colourless gum. Both 1H- and 13C- dried flowers and leaves of U. hecatantha (300 g) were NMR data of santhemoidin C (3) in DMSO-d6 as a solvent extracted thrice at room temperature with dichloromethane are reported here: δ 169.9 (C-1’’; acetate carbonyl), 169.0 (4.5 L, 6 h). Filtrates were joined and concentrated on a (C-12), 164.5 (C-1′), 150.2 (C-3′), 144.2 (C-4), 136.9 (C- rotary evaporator at 40°C under reduced pressure to give 11), 133.5 (C-10), 129.1 (C-1), 125.2 (C-2′), 123.0 (C-5), 42 g of crude extract (DE). *e crude extract was suspended 120.7 (C-13), 76.6 (C-3), 75.0 (C-6), 72.6 (C-8), 57.7 in ethanol (147 mL) at 60°C, diluted with distilled water (C-4′), 57.5 (C-5′), 50.8 (C-7), 42.8 (C-9), 35.4 (C-2), 20.2 (63 mL), and filtered under vacuum. *e filtrate was (C-2’’; acetate methyl), 18.6 (C-14), and 11.8 ppm (C-15); extracted successively with hexane (3 × 60 mL) and 1H-NMR data for santhemoidin C in DMSO-d-6 at Evidence-Based Complementary and Alternative Medicine 3

600 MHz: δ 4.97 dd (12 and 3.5 Hz; H-1), 2.16 ddd (12, 12, in an axenic medium (BHI-Tryptose supplemented with: 10; H-2α), 2.30 m (H-2β), 4.15 ddd (10, 5.5, 4.5; H-3), FBS 10%, hemine 2 ×10−5 mg/mL, glucose 3.0 ×10−4 g/mL, 4.91 d br (9.8; H-5), 5.08 dd (9.8, 8.5; H-6), 3.22 ddd (8.5, streptomycin 2.0 ×10−4 g/mL, ampicillin 1.3 ×10−4 g/mL) as 3.5, 3; H-7), 5.71 m (H-8), 2.64 dd (14.2, 4.4; H-9a), 2.40 dd a low-cost alternative for Leishmania spp. culture. Assays (14.2, 2; H-9b), 6.14 d (3.5; H-13a), 5.65 d (3.0; H-13b), were performed in 96-well plastic plates using 2 ×106 pro- 1.40 s br (3H; H-14), 1.67 s br (3H; H-15), 6.97 t (5.7; mastigotes per well. Compounds were dissolved in DMSO. H-3′), 4.27 t (2H; 5.7; H-4′), 4.76 d (12.1; 5′a), 4.71 d (12.1; Different serial dilutions (25–0.05 µM) of the compounds 5′b), 1.91 s (3H; C-2’’; acetate methyl); Others: O–H at with a final volume up to 200 µL were added. After 48 h at C-3, 5.29 d (4.5); O–H at C-4′, 5.18 t (5.7). 28°C, 20 µL of a 2 mM resazurin solution was added, and the See spectra in supplementary material. Assignments oxidation-reduction was quantified at 570 and 600 nm. *e were made by H–H COSY, HSQC, and HMBC experiments. resazurin solution was prepared at 2.5 mM in phosphate- buffered solution (PBS), pH 7.4, and filtered through 0.22 µm before use. Resazurin sodium salt was obtained 2.4. Spectrometric Analyses. *e isolated compounds were from Sigma-Aldrich (St. Louis, MO, USA) and stored at 4°C identified by proton nuclear magnetic resonance (1H- protected from light. Glucantime was used as a positive NMR) and carbon nuclear magnetic resonance (13C- control. *e efficacy of each compound was estimated by NMR), heteronuclear single quantum correlation calculating the IC values using OriginLab 8.5® sigmoidal (HSQC), heteronuclear multiple bond correlation 50 regression. Each antiproliferative experiment was done in (HMBC), correlated spectroscopy (COSY) (Bruker Ad- duplicate, and each concentration was tested in triplicate. vance 600) (600 MHz in CDCl3), and electron impact- mass spectrometry (EI-MS). *e purity of santhemoidin C (3), estimated by 1H- 2.6. Cytotoxicity Assay. *e cytotoxicity of the dewaxed NMR, was >95%. Likewise, the purity estimated for euparin dichloromethane extract of U. hecatantha and the iso- (1) and eucannabinolide (4), also by 1H-NMR, was 97.5% lated compounds was evaluated according to the method and ca. 94%, respectively (see the corresponding 1H-NMR described by Aguilera et al., 2019 [15]. *e J774.1 murine spectra in supplementary material). Jaceidin sample was macrophages (ATCC, USA) were grown in a DMEM analyzed by TLC using CH2Cl2 : EtOAc as a solvent and a culture medium containing 4 mM L-glutamine and sup- 10% solution of antimony (III) chloride in chloroform as plemented with 10% FCS. Cells were seeded in a 96-well spray reagent. A single spot was observed under long-wave plate (5.00 ×104 cells in 200 µL culture medium) and ° ° UV light. It was identified by its mp 131–135 C (“Jaceidin,” incubated at 37 C in a 5% CO2 atmosphere for 48 h, to Human Metabolome Database, HMDB0033819) and by allow cell adhesion before drug testing. Afterwards, cells chromatographic analysis with an authentic sample and were exposed for 48 h to the compounds (25–400 µM) or confirmed by UV spectroscopy with shift reagents. the vehicle for control (medium with 0.4% DMSO v/v), and additional control (cells in medium) were used in each test. Cell viability was then assessed by measuring 2.5. Antiparasitic Activity Assay. For the anti-Trypanosoma the mitochondria-dependent reduction of MTT [3-(4,5- and anti-Leishmania activity assays, we followed the dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] methods described by Aguilera et al., 2019 [15]. to formazan. For this purpose, MTT in sterile PBS For the in vitro anti-Trypanosoma cruzi activity, (containing 0.2% glucose), pH 7.4, was added to the epimastigotes of the Tulahuen 2 strain (genotype TcVI) macrophages to achieve a final concentration of 0.1 mg/ grown in an axenic medium (BHI-Tryptose) were used. mL, and the cells were incubated at 37°C for 3 h. After Cells from a 5–7-day-old culture were inoculated in a removing the medium, formazan crystals were dissolved fresh culture medium to give an initial concentration of in 180 µL of DMSO and 20 µL of MTT buffer (0.1 M 106 cells/mL. *e absorbance at 600 nm of the cells in glycine, 0.1 M NaCl, 0.5 mM EDTA, pH 10.5), and the culture was measured every day. On day five, the medium absorbance at 560 nm was measured. *e CC was de- was inoculated with different doses of the compounds 50 fined as the drug concentration at which 50% of the cells (25–0.05 µM) from a stock solution in dimethylsulfoxide were viable, relative to the control (no drug added), and (DMSO) (DMSO concentration in the culture medium was determined using OriginLab 8.5 sigmoidal re- never exceeded 0.4%). Control parasites were cultivated gression (% of viable cells compared to® the logarithm of in medium with 0.4% DMSO v/v. Benznidazole was used the compound concentration). Tests were performed in as a positive control. At five days, the absorbance of the triplicate. culture was measured and compared to the control and the IC50 values were calculated for each compound using OriginLab 8.5 sigmoidal regression. Each experiment 2.7. Toxicity and Pharmacokinetic Properties. *e toxicity was done in duplicate,® and each concentration was tested and pharmacokinetic properties of the compounds were in triplicate. estimated with the open-access SwissADME software Leishmania infantum (MHOM/BR/2002/LPC-RPV) (http://www.swissadme.ch), a tool that allows the pre- was obtained from Fiocruz (Collection of Oswaldo Cruz diction of different pharmacokinetic parameters such as Foundation, , ). Promastigotes were water solubility, gastrointestinal absorption, skin pene- cultured as described [16] with some modifications at 28°C trability, lipophilicity, bioavailability, and so forth and 4 Evidence-Based Complementary and Alternative Medicine

T. E. S. T (Toxicity Estimation Software Tool). *e *e cytotoxic effect of compounds 1–4 was assayed on software input uses the SMILES codes of the molecules, mammalian cells. *e terpenoid compounds, santhe- which were generated with the ChemBioOffice 2010 moidin C (3) and eucannabinolide (4), showed CC50 program. values of >15 µM and 15 µM, respectively. Euparin (1) and jaceidin (2) presented CC50 values > 25 µM. Taking in consideration that selectivity is a relevant characteristic 2.8. Statistical Analysis. *e statistical analysis was per- for defining hit molecules, selectivity indexes (SI) of the formed using Origin software package version 7.0. *e compounds were calculated. *e most active compound statistical significance of the difference between the data against T. cruzi, eucannabinolide (4), showed a SI value of pairs was evaluated by analysis of variance (one-way 1.5. ANOVA), followed by the Tukey test. Statistical differences *e differences in activity between T. cruzi and were considered significant at p < 0.05. L. infantum for compound 4 compared to compound 3 are remarkable. Both are germacranolides with the same 3. Results and Discussion molecular formula (C22H28O8) but differ in the stereo- chemistry of the C4–C5 double bond: lactone 3 is a *e dewaxed dichloromethane extract (DDE) of the aerial germacrolide (a trans, trans-germacranolide) while lac- parts of U. hecatantha was evaluated against T. cruzi tone 4 is a heliangolide (a trans, cis-germacranolide) [22]; epimastigotes and L. infantum promastigotes. *is extract they also differ in the location of the acetyl group which was active against T. cruzi and L. infantum with 50% in- in lactone 3 esterifies the hydroxyl group of the 4,5- hibitory concentration (IC50) values of 7 µg/mL and 31 µg/ dihydroxytigloyloxy ester residue at C-8, while, in lactone mL, respectively. *e in vitro cytotoxic effect of the DDE 4, it esterifies the hydroxyl group at C-3 of the heli- was evaluated on murine macrophages by the MMT angolide skeleton. *ese differences should be expected method. *is extract showed a 50% cytotoxic concentration to strongly affect conformation, electronic distribution, (CC50) value of 15 µg/mL. Fractionation of the DDE by and hydrogen bonding interactions. *erefore, these column chromatography and purification of the sub- stereochemical and positional changes could increase the fractions by chromatographic techniques yielded four activity of compound 4 on T. cruzi and decrease it for compounds: compound 1 (0.0093%), compound 2 L. infantum. Santhemoidin C, euparin, and jaceidin did (0.0033%), compound 3 (0.04%), and compound 4 not display selectivity on T. cruzi epimastigotes. None of (0.036%). *e compounds were identified by spectroscopic the compounds showed selectivity against L. infantum methods as euparin (1), jaceidin (2), santhemoidin C (3), promastigotes. and eucannabinolide (4) (Figure 1). *e pharmacokinetic characteristics and toxicity of the Euparin and eucannabinolide have been previously compounds play an important role in the drug discovery isolated from U. hecatantha collected in Bolivia [14]. *e process. *ese properties are influenced in part by the presence of santhemoidin C and jaceidin in this species is physicochemical properties of drugs. In this sense, the reported for the first time. *e sesquiterpene lactones mutagenicity, the oral rat LD50, the Log P, solubility, gas- santhemoidin C and eucannabinolide have been described trointestinal (GI) absorption, skin permeation (Log Kp), and in Schkuhria anthemoidea [17]. Eucannabinolide, jaceidin, the blood-brain barrier (BBB) permeant were predicted and euparin have also been reported in other Asteraceae (Table 1). Euparin (1), santhemoidin C (3), and eucanna- species [18–21]. binolide (4) showed no mutagenicity and LD50 val- *e in vitro antiprotozoal activity of the isolated ues > 600 mg/kg with high GI absorption. Santhemoidin C compounds (1–4) was evaluated against T. cruzi epi- and eucannabinolide presented log P values lower than 2, mastigotes and L. infantum promastigotes (Figure 2). *e satisfying the criteria established by Lipinski [23] and sesquiterpene lactone eucannabinolide (4) was the most showed the best skin permeation. Compounds 2–4 showed active on T. cruzi with an IC50 value of 10 ± 2 µM (4.2 µg/ no BBB permeation. mL). Santhemoidin C (3) displayed also a trypanocidal Eucannabinolide has shown activity against Trypanosoma activity with an IC50 of 18 ± 3 µM (7.6 µg/mL). On the brucei rhodesiense trypomastigotes (IC50 �1.1 ± 0.1 µM) and other hand, euparin (1) and jaceidin (2) showed moderate has tested its cytotoxicity on mammalian cells (L6-cell line activity against epimastigotes with IC50 values > 25 µM. from rat-skeletal myoblasts; CC50 � 7.8 ± 0.8 µM) [24]. *is *e IC50 for the positive control benznidazole was sesquiterpene lactone was also active when it was loaded onto 7 ± 2 µM. polylactic acid nanoparticles with a free drug equivalent Euparin (1) and santhemoidin C (3) were the most active IC50 value of 3.32 µM [25]. Eucannabinolide has also on L. infantum with IC50 values of 18 ± 4 µM (3.9 µg/mL) exhibited cytotoxic activity against tumour cell lines [26] and and 19 ± 4 µM (8.0 µg/mL), respectively. *e flavonoid anti-inflammatory activity [27]. Compound 4 has also shown jaceidin (2) and the sesquiterpene lactone eucannabinolide drug-like pharmacokinetic and toxicity properties (Table 1). (4) were less active (IC50 > 25 µM). Glucantime showed an No reports about the biological activities of santhemoidin C IC50 value of 26 ± 9 µM. have been found. Antibacterial and antiviral activity [28, 29] Evidence-Based Complementary and Alternative Medicine 5

O

O OH

O HO HO O

O O

OH O

(a) (b) OH OH

O O

O OAc O OH O

HO O

O O

O O

(c) (d)

Figure 1: Chemical structures of euparin (a), jaceidin (b), santhemoidin C (c), and eucannabinolide (d).

120

100

80

60

40 Growth inhibition (%)

20

0 1 234 T. cruzi L. infantum

Figure 2: Effect of compounds 1–4 (25 µM) on T. cruzi and L. infantum. 6 Evidence-Based Complementary and Alternative Medicine

Table 1: Predicted toxicity and pharmacokinetic profile of the isolated compounds. Mutagenicity by Oral rat LD Consensus Solubility (mg/ GI Log kp (skin BBB Compound 50 Ames test (mg/kg) Log P o/w mL) absorption permeation) cm/s permeant 1 Negative 620 2.82 3.77e − 02 High −5.03 Yes 2 Positive 303 2.15 3.42e − 02 High −6.52 No 3 Negative 680 1.69 1.85 High −8.45 No 4 Negative 680 1.63 1.85 High −8.45 No and protective effect on human lymphocytes against chro- Investigacion´ Cient´ıfica, Programa Grupos I+D, de la mosomal damage [30] have been reported for jaceidin. Universidad de la Republica,´ Uruguay. Euparin has shown antipoliovirus activity [31] and cytotoxic activity against liver carcinoma cells [32]. *is is the first time that the activity against T. cruzi and L. infantum of these Supplementary Materials compounds has been reported. *e NMR and MS spectra of the compounds were included as supplementary materials. (Supplementary Materials) 4. Conclusions In this study, the activity against T. cruzi and L. infantum of References the dichloromethane extract of U. hecatantha and the iso- [1] World Health Organization (WHO), Neglected Tropical lation of four compounds, euparin, jaceidin, santhemoidin Diseases, World Health Organization (WHO), Geneva, C, and eucannabinolide, are reported. *is is the first Switzerland, 2020, https://www.who.int/neglected_diseases/ communication describing the isolation of santhemoidin C diseases/en/. and jaceidin from U. hecatantha. *e activity of the isolated [2] World Health Organization (WHO), Chagas Disease compounds against T. cruzi and L. infantum is being re- (American Trypanosomiasis), World Health Organization ported for the first time. *e sesquiterpene lactone eucan- (WHO), Geneva, Switzerland, 2020, https://www.who.int/ nabinolide was the most active compound against T. cruzi health-topics/chagas-disease#tab=tab_1. and could be considered for further studies. [3] K. C. F. Lidani, F. A. Andrade, L. Bavia et al., “Chagas disease: from discovery to a worldwide health problem,” Frontiers in Public Health, vol. 7, 166 pages, 2019. Data Availability [4] J. A. Perez-Molina,´ C. Crespillo-And´ujar,P. Bosch-Nicolau, and I. Molina, “Trypanocidal treatment of Chagas disease,” *e data used to support the findings of this study are in- Enfermedades Infecciosas Y Microbiolog´ıa Cl´ınica, vol. 20, cluded within the article. pp. 30193–30202, 2020. [5] World Health Organization (WHO), Leishmaniasis, World Disclosure Health Organization (WHO), Geneva, Switzerland, 2020, https://www.who.int/health-topics/leishmaniasis#tab=tab_1. *is investigation is part of the activities carried out within [6] A. Bhattacharya, A. Corbeil, R. L. do Monte-Neto et al., “Of the “Research Network Natural Products against Neglected drugs and trypanosomatids: new tools and knowledge to Diseases” (ResNet NPND): http://www.resnetnpnd.org/. reduce bottlenecks in drug discovery,” Genes (Basel), vol. 11, no. 7, p. 722, 2020. [7] Y. Tu, “Artemisinin-A gift from traditional Chinese medicine Conflicts of Interest to the World (nobel lecture),” Angewandte Chemie Interna- tional Edition, vol. 55, no. 35, pp. 10210–10226, 2016. *e authors declare that there are no conflicts of interest [8] N. Tajuddeen and F. R. Van Heerden, “Antiplasmodial natural regarding the publication of this paper. products: an update,” Malaria Journal, vol. 18, no. 1, p. 404, 2019. Authors’ Contributions [9] T. J. Schmidt, S. A. Khalid, A. J. Romanha et al., “*e potential of secondary metabolites from as drugs or leads against *e work was codirected by Guzman Alvarez and Valeria protozoan neglected diseases - part II,” Current Medicinal P. Sulsen.¨ Orlando G. Elso and Maria Clavin contributed Chemistry, vol. 19, no. 14, pp. 2176–2228, 2012. equally to this work. [10] R. M. King and H. Robinson, “*e genera of the (Asteraceae),” Monographs in Systematic Botany, vol. 22, pp. 89–92, 1987. Acknowledgments [11] A. Filipov, “Medicinal plants of the pilaga´ of central chaco,” Journal of Ethnopharmacology, vol. 44, no. 3, pp. 181–193, *is work was supported by Grant nos. 11220150100158CO, 1994. PICT 2015–3531, and UBACYT 20020170100316BA from [12] G. J. Mart´ınez and G. E. Barboza, “Natural pharmacopoeia the National Scientific and Technical Research Council, the used in traditional Toba medicine for the treatment of par- National Agency for Science and Technology Promotion, asitosis and skin disorders (Central Chaco, Argentina),” and the University of Buenos Aires. *is work was sup- Journal of Ethnopharmacology, vol. 132, no. 1, pp. 86–100, ported in part by Grant no. 35 from Comision´ Sectorial de 2010. Evidence-Based Complementary and Alternative Medicine 7

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